Novel composition for preparing polysaccharide fibers

ABSTRACT

Solutions formed by combining poly(α(1→3) glucan) with CS 2  in aqueous alkali metal hydroxide solution have been shown to produce the xanthated form of the poly(α(1→3) glucan). The solutions so formed have been shown to be useful for solution spinning into fiber of poly(α(1→3) glucan) when the spun fiber is coagulated in an acidic coagulation bath. The fibers so produced exhibit desirable physical properties. The poly(α(1→3) glucan) employed was synthesized by fermentation.

FIELD OF THE INVENTION

The present invention is directed to a process for forming fibers ofpoly(α(1→3) glucan) by solution spinning a solution of xanthatedpoly(α(1→3) glucan) in an aqueous alkali metal hydroxide and to thesolution itself. The poly(α(1→3) glucan) employed was synthesized byfermentation.

BACKGROUND OF THE INVENTION

Polysaccharides have been known since the dawn of civilization,primarily in the form of cellulose, a polymer formed from glucose bynatural processes via β(1→4) glycoside linkages; see, for example,Applied Fibre Science, F. Happey, Ed., Chapter 8, E. Atkins, AcademicPress, New York, 1979. Numerous other polysaccharide polymers are alsodisclosed therein.

Only cellulose among the many known polysaccharides has achievedcommercial prominence as a fiber. In particular, cotton, a highly pureform of naturally occurring cellulose, is well-known for its beneficialattributes in textile applications.

It is further known that cellulose exhibits sufficient chain extensionand backbone rigidity in solution to form liquid crystalline solutions;see, for example O'Brien, U.S. Pat. No. 4,501,886. The teachings of theart suggest that sufficient polysaccharide chain extension could beachieved only in β(1→4) linked polysaccharides and that any significantdeviation from that backbone geometry would lower the molecular aspectratio below that required for the formation of an ordered phase.

More recently, glucan polymer, characterized by α(1→3) glycosidelinkages, has been isolated by contacting an aqueous solution of sucrosewith GtfJ glucosyltransferase isolated from Streptococcus salivarius,Simpson et al., Microbiology, vol 141, pp. 1451-1460 (1995). Highlycrystalline, highly oriented, low molecular weight films ofα(1→3)-D-glucan have been fabricated for the purposes of x-raydiffraction analysis, Ogawa et al., Fiber Diffraction Methods, 47, pp.353-362 (1980). In Ogawa, the insoluble glucan polymer is acetylated,the acetylated glucan dissolved to form a 5% solution in chloroform andthe solution cast into a film. The film is then subjected to stretchingin glycerine at 150° C. which orients the film and stretches it to alength 6.5 times the original length of the solution cast film. Afterstretching, the film is deacetylated and crystallized by annealing insuperheated water at 140° C. in a pressure vessel. It is well-known inthe art that exposure of polysaccharides to such a hot aqueousenvironment results in chain cleavage and loss of molecular weight, withconcomitant degradation of mechanical properties.

Polysaccharides based on glucose and glucose itself are particularlyimportant because of their prominent role in photosynthesis andmetabolic processes. Cellulose and starch, both based on molecularchains of polyanhydroglucose are the most abundant polymers on earth andare of great commercial importance. Such polymers offer materials thatare environmentally benign throughout their entire life cycle and areconstructed from renewable energy and raw materials sources.

The term “glucan” is a term of art that refers to a polysaccharidecomprising beta-D-glucose monomer units that are linked in eightpossible ways. Cellulose is a glucan.

Within a glucan polymer, the repeating monomeric units can be linked ina variety of configurations following an enchainment pattern. The natureof the enchainment pattern depends, in part, on how the ring closes whenan aldohexose ring closes to form a hemiacetal. The open chain form ofglucose (an aldohexose) has four asymmetric centers (see below). Hencethere are 2⁴ or 16 possible open chain forms of which D and L glucoseare two. When the ring is closed, a new asymmetric center is created atC1 thus making 5 asymmetric carbons. Depending on how the ring closes,for glucose, α(1→4)-linked polymer, e.g. starch, or β(1→4)-linkedpolymer, e.g. cellulose, can be formed upon further condensation topolymer. The configuration at C1 in the polymer determines whether it isan alpha or beta linked polymer, and the numbers in parenthesisfollowing alpha or beta refer to the carbon atoms through whichenchainment takes place.

The properties exhibited by a glucan polymer are determined by theenchainment pattern. For example, the very different properties ofcellulose and starch are determined by the respective nature of theirenchainment patterns. Starch or amylose consists of α(1→4) linkedglucose and does not form fibers among other things because it isswollen or dissolved by water. On the other hand, cellulose consists ofβ(1→4) linked glucose, and makes an excellent structural material beingboth crystalline and hydrophobic, and is commonly used for textileapplications as cotton fiber, as well as for structures in the form ofwood.

Like other natural fibers, cotton has evolved under constraints whereinthe polysaccharide structure and physical properties have not beenoptimized for textile uses. In particular, cotton fiber is of shortfiber length, limited variation in cross section and fiber fineness andis produced in a highly labor and land intensive process.

O'Brien, U.S. Pat. No. 7,000,000 discloses a process for preparing fiberfrom liquid crystalline solutions of acetylated poly(α(1→3) glucan). Thethus prepared fiber was then de-acetylated resulting in a fiber ofpoly(α(1→3) glucan).

SUMMARY OF THE INVENTION

Considerable benefit accrues to the process hereof that provides ahighly oriented and crystalline poly(α(1→3) glucan) fiber withoutsacrifice of molecular weight by the solution spinning of fiber from thenovel solution hereof.

In one aspect the present invention is directed to a solution comprising0.75 to 2 molar aqueous alkali metal hydroxide and a solids content of 5to 20% by weight of xanthated poly(α(1→3) glucan); wherein the numberaverage molecular weight of the xanthated poly(α(1→3) glucan) is atleast 10,000 Daltons; and, wherein the degree of xanthation of thexanthated poly(α(1→3) glucan) lies in the range of 0.1 to 1.

In another aspect, the present invention is directed to a processcomprising forming a solution by dissolving in a 0.75 to 2 molar aqueousalkali metal hydroxide, CS₂, and 5 to 20 percent by weight of the totalweight of the resulting solution of poly(α(1→3) glucan) characterized bya number average molecular weight of at least 10,000 Da, causing saidsolution to flow through a spinneret, forming a fiber thereby; andcontacting said fiber with an acidic liquid coagulant; wherein saidprocess the weight ratio of CS₂ to poly(α(1→3) glucan) lies in the rangeof 0.1 to 1.0.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a schematic diagram of an apparatus suitable for air gap orwet spinning of the aqueous alkali metal hydroxide solutions of PAGXhereof.

DETAILED DESCRIPTION

When a range of values is provided herein, it is intended to encompassthe end-points of the range unless specifically stated otherwise.Numerical values used herein have the precision of the number ofsignificant figures provided, following the standard protocol inchemistry for significant figures as outlined in ASTM E29-08 Section 6.For example, the number 40 encompasses a range from 35.0 to 44.9,whereas the number 40.0 encompasses a range from 39.50 to 40.49.

The term “solids content” is a term of art. It is used herein to referto the percentage by weight of xanthated poly(α(1→3) glucan) (PAGX) inthe aqueous alkali metal hydroxide solution hereof (MOH (aq). It iscalculated from the formula:

${SC} = {\frac{{Wt}({PAGX})}{{{Wt}({PAGX})} + {{Wt}\left( {{MOH}({aq})} \right)}} \times 100}$

where SC represents “solids content,” and Wt(PAGX), Wt(MOH(aq)) arerespectively weights of the poly(α(1→3) glucan) xanthate (PAGX), and ofthe aqueous alkali metal hydroxide. The term “solids content” issynonymous with the concentration by weight of xanthated poly(α(1→3)glucan) with respect to the total weight of solution.

Percent by weight is represented by the term “wt-%.”

The formula “MOH” shall be employed to refer to the alkali metalhydroxide suitable for the practice of the invention. The formula“MOH(aq)” shall be employed to refer to the aqueous alkali metalhydroxide solution suitable for the practice of the invention. It shallbe understood that the expression “concentration of the MOH(aq)” shallrefer to the molarity of the aqueous alkali metal hydroxide solutionhereof.

A polymer, including glucan, and poly(α(1→3) glucan) (PAG) inparticular, is made up of a plurality of so-called repeat unitscovalently linked to one another. The repeat units in a polymer chainare diradicals, the radical form providing the chemical bonding betweenrepeat units. For the purposes of the present invention the term“glucose repeat units” shall refer to the diradical form of glucose thatis linked to other diradicals in the polymer chain, thereby forming saidpolymer chain.

The term “glucan” refers to polymers, including oligomers and lowmolecular weight polymers that are unsuitable for fiber formation. Forthe purposes of the present invention, the glucan polymer suitable forthe practice of the invention is a poly(α(1→3) glucan) or xanthatedpoly(α(1→3) glucan), characterized by a number average molecular weightof at least 10,000 Daltons, preferably of at least 40,000-100,000Daltons.

Suitable PAGX is characterized by a degree of xanthation in the range of0.1 to 1. The term “xanthation” is a term of art referring to thereaction of a hydroxyl group with CS₂ in alkali metal hydroxide,according to the following reaction:

In the case of the PAG suitable for use in the process of the invention,each cyclic hexose repeat unit offers three hydroxyls for potentialreaction to form the xanthate according to the above reaction scheme.The term “degree of xanthation” refers to the average percentage ofavailable hydroxyl sites in each repeat unit that have actuallyundergone reaction to the xanthate. The theoretical maximum degree ofxanthation a suitable PAG polymer molecule can undergo is 3—that is,every single hydroxyl site in the polymer would have undergone reaction.

According to the present invention, suitable PAGX polymers haveundergone xanthation to the degree of 0.1 to 1. This means that on theaverage between one hydroxyl site per ten repeat units, and 10 hydroxylsites per ten repeat units have undergone the xanthation reaction, whilethe theoretical maximum would be 30 hydroxyl sites per ten repeat units.

In one aspect the present invention is directed to a solution comprising0.75 to 2 molar aqueous alkali metal hydroxide and a solids content of 5to 20% by weight of PAGX; wherein the number average molecular weight ofthe PAGX is at least 10,000 Daltons; and, wherein the degress ofxanthation of the PAGX lies in the range of 0.1 to 1.

In one embodiment, the alkali metal hydroxide (MOH) is sodium hydroxide.In a further embodiment the concentration of the NaOH is in the range of1.0 to 1.7 M.

In one embodiment, the solids concentration is in the range of 7.5 to15%.

The PAG suitable for use in the process of the present invention is aglucan characterized by a number average molecular weight (M_(n)) of atleast 10,000 Da wherein at least 90 mol-% of the repeat units in thepolymer are glucose repeat units and at least 50% of the linkagesbetween glucose repeat units are α(1→3) glycoside linkages. Preferablyat least 95 mol-%, most preferably 100 mol-%, of the repeat units areglucose repeat units. Preferably at least 90%, most preferably 100%, ofthe linkages between glucose units are α(1→3) glycoside linkages.

The isolation and purification of various polysaccharides is describedin, for example, The Polysaccharides, G. O. Aspinall, Vol. 1, Chap. 2,Academic Press, New York, 1983. Any means for producing the α(1→3)polysaccharide suitable for the invention in satisfactory yield and 90%purity is suitable. In one such method, disclosed in U.S. Pat. No.7,000,000, poly(α(1→3)-D-glucose) is formed by contacting an aqueoussolution of sucrose with gtfJ glucosyltransferase isolated fromStreptococcus salivarius according to the methods taught in the art. Inan alternative such method, the gtfJ is generated by geneticallymodified E. Coli, as described in detail, infra.

The PAG suitable for use in the present invention can further compriserepeat units linked by a glycoside linkage other than α(1→3), includingα(1→4), α(1→6), β(1→2), β(1→3), β(1→4) or β(1→6) or any combinationthereof. According to the present invention, at least 50% of theglycoside linkages in the polymer are α(1→3) glycoside linkages.Preferably at least 90%, most preferably 100%, of the linkages betweenglucose units are α(1→3) glycoside linkages.

The solution hereof is prepared by adding a suitable PAG to MOH(aq),containing carbon disulfide and agitating to obtain thorough mixing.PAGX is formed in situ under these conditions. The solids content ofPAGX in the solution ranges from 5 to 20% by weight with respect to thetotal weight of the solution. When solids content of PAGX is below 5%,the fiber-forming capability of the solution is greatly degraded.Solutions with solids content above 15% are increasingly problematicalto form, requiring increasingly refined solution forming techniques.

In any given embodiment, the solubility limit of PAGX is a function ofthe molecular weight of the PAGX, the concentration of the MOH(aq), thedegree of xanthation, the duration of mixing, the viscosity of thesolution as it is being formed, the shear forces to which the solutionis subject, and the temperature at which mixing takes place. Generally,higher shear mixing and higher temperature will be associated withhigher solubility. The maximum temperature for mixing is limited 46° C.,the boiling point of the CS₂. From the standpoint of solubility andspinnability, the optimum concentrations of the MOH(aq) and CS₂ maychange depending upon the other parameters in the mixing process.

In the practice of the invention, it has been observed that the reactionof the CS₂ with the PAG to form the xanthate occurs quantitativelywithin about one to three hours at room temperature. The xanthate soformed has also been observed to be chemically unstable, degradingcompletely into a variety of by-products after approximately 36 hours ofsolution time. It is therefore incumbent upon the practitioner hereof toemploy the solution hereof for fiber spinning after the time requiredfor formation of the xanthate but before significant degradation canoccur. For a solution hereof prepared at room temperature, spinning isthus performed preferably between 1 to 3 hours of solution time,depending upon the reaction time for xanthate formation. The term“solution time” refers to the time elapsed from when the ingredients ofthe solution are first combined. Thus, in a preferred embodiment of theprocess hereof, the ingredients are combined, allowed to stand for 1 to3 hours, and then spun into fiber as described in detail, infra. In asomewhat less preferred embodiment chemically, but more preferred from apractical viewpoint, a solution time on the order of 1-5 hours is alsosuitable.

The present invention is further directed to a process comprisingforming a solution by dissolving in a 0.75 to 2 molar aqueous alkalimetal hydroxide, CS₂, and 5 to 15 percent by weight of the total weightof the resulting solution of PAG characterized by a number averagemolecular weight of at least 10,000 Da; causing said solution to flowthrough a spinneret, forming a fiber thereby; and, contacting said fiberwith an acidic liquid coagulant; wherein said process the weight ratioof CS₂ to PAG lies in the range of 0.1 to 1.0.

In one embodiment, the alkali metal (M) is sodium.

In a further embodiment of the process hereof, a suitable PAG is onewherein 100% of the repeat units are glucose, and 100% of the linkagesbetween glucose repeat units are α(1→3) glycoside linkages.

In the process hereof, the minimum solids content of PAGX required inthe solution in order to achieve stable fiber formation varies accordingto the molecular weight of the PAGX, as well as the degree ofxanthation. It is found in the practice of the invention that a 5%solids content is an approximate lower limit to the concentration neededfor stable fiber formation. At >15%, especially greater than 20% solids,excessive amounts of undissolved PAGX are present, causing a degradationin fiber spinning performance. A solution having a solids content of atleast 7.5% is preferred. A solids content ranging from about 7.5% toabout 15% in a 1.0 to 1.7 M NaOH solution is more preferred. Preferredis a PAGX characterized by a number average molecular weight in therange of 40,000-100,000 Daltons and degree of xanthation in the range of0.1-1.

Spinning from the solution hereof can be accomplished by means known inthe art, and as described in O'Brien, op. cit. The viscous spinningsolution can be forced by means such as the push of a piston or theaction of a pump through a single or multi-holed spinneret or other formof die. The spinneret holes can be of any cross-sectional shape,including round, flat, multi-lobal, and the like, as are known in theart. The extruded strand can then be passed by ordinary means into acoagulation bath wherein is contained a liquid coagulant which convertsthe PAGX back to PAG, causing the polymer to coagulate into a fiberaccording to the present invention.

Suitable liquid coagulants include but are not limited to glacial aceticacid, aqueous acetic acid, sulfuric acid, combinations of sulfuric acid,sodium sulfate, and zinc sulfate. In one embodiment, the liquidcoagulant is maintained at a temperature in the range of 0-100° C., andpreferably in the range of 15-70° C.

In one embodiment, the coagulation bath comprises glacial acetic acid.It is found in the practice of the invention that satisfactory resultsare achieved by employing as the coagulant liquid an excess of glacialacetic acid. During the course of spinning, the glacial acetic acidneutralizes aqueous NaOH and regenerates PAG from PAGX as the spun fiberpasses through the coagulant bath.

In a preferred embodiment, extrusion is effected directly into thecoagulation bath. In such a circumstance, known in the art as“wet-spinning,” the spinneret is partially or fully immersed in thecoagulation bath. The spinnerets and associated fittings should beconstructed of corrosion resistant alloys such as stainless steel orplatinum/gold.

In one embodiment, the thus coagulated fiber is then passed into asecond bath provided to neutralize and dilute residual acid from thecoagulation bath. The secondary bath preferably contains H₂O, methanol,or 5% aqueous NaHCO₃, or a mixture thereof. Aqueous NaHCO₃ is preferred.In an embodiment, the wound fiber package is soaked in one or moreneutralizing wash baths for a period of time up to four hours in eachbath. A sequence of baths comprising respectively 5% aqueous NaHCO₃,methanol, and H₂O, has been found satisfactory.

The invention hereof is further described in, but not limited by, thefollowing specific embodiments thereof.

EXAMPLES Preparation of Glucosyltransferase (GtfJ) Enzyme Materials

Dialysis tubing (Spectrapor 25225-226, 12000 molecular weight cut-off)was obtained from VWR (Radnor, Pa.).

Dextran and ethanol were obtained from Sigma Aldrich. Sucrose wasobtained from VWR.

Suppressor 7153 antifoam was obtained from Cognis Corporation(Cincinnati, Ohio).

All other chemicals were obtained from commonly used suppliers.

Seed Medium

The seed medium, used to grow the starter cultures for the fermenters,contained: yeast extract (Amberx 695, 5.0 grams per liter (g/L)), K₂HPO₄(10.0 g/L), KH₂PO₄ (7.0 g/L), sodium citrate dihydrate (1.0 g/L),(NH₄)₂SO₄ (4.0 g/L), MgSO₄ heptahydrate (1.0 g/L) and ferric ammoniumcitrate (0.10 g/L). The pH of the medium was adjusted to 6.8 usingeither 5N NaOH or H₂SO₄ and the medium was sterilized in the flask. Poststerilization additions included glucose (20 mL/L of a 50% w/w solution)and ampicillin (4 mL/L of a 25 mg/mL stock solution).

Fermenter Medium

The growth medium used in the fermenter contained: KH₂PO₄ (3.50 g/L),FeSO₄ heptahydrate (0.05 g/L), MgSO₄ heptahydrate (2.0 g/L), sodiumcitrate dihydrate (1.90 g/L), yeast extract (Ambrex 695, 5.0 g/L),Suppressor 7153 antifoam (0.25 milliliters per liter, mL/L), NaCl (1.0g/L), CaCl₂ dihydrate (10 g/L), and NIT trace elements solution (10mL/L). The NIT trace elements solution contained citric acid monohydrate(10 g/L), MnSO₄ hydrate (2 g/L), NaCl (2 g/L), FeSO₄ heptahydrate (0.5g/L), ZnSO₄ heptahydrate (0.2 g/L), CuSO₄ pentahydrate (0.02 g/L) andNaMoO₄ dihydrate (0.02 g/L). Post sterilization additions includedglucose (12.5 g/L of a 50% w/w solution) and ampicillin (4 mL/L of a 25mg/mL stock solution).

Construction of Glucosyltransferase (gtfJ) Enzyme Expression Strain

A gene encoding the mature glucosyltransferase enzyme (GtfJ; EC 2.4.1.5;GENBANK® AAA26896.1, SEQ ID NO: 3) from Streptococcus salivarius (ATCC25975) was synthesized using codons optimized for expression in E. coli(DNA 2.0, Menlo Park Calif.). The nucleic acid product (SEQ ID NO: 1)was subcloned into pJexpress404® (DNA 2.0, Menlo Park Calif.) togenerate the plasmid identified as pMP52 (SEQ ID NO: 2). The plasmidpMP52 was used to transform E. coli MG1655 (ATCC 47076) to generate thestrain identified as MG1655/pMP52.

Standard recombinant DNA and molecular cloning techniques used hereinare well known in the art and are described by Sambrook, J. and Russell,D., Molecular Cloning: A Laboratory Manual, Third Edition, Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y. (2001); and bySilhavy, T. J., Bennan, M. L. and Enquist, L. W., Experiments with GeneFusions, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY(1984); and by Ausubel, F. M. et. al., Short Protocols in MolecularBiology, 5^(th) Ed. Current Protocols, John Wiley and Sons, Inc., N.Y.,2002.

Materials and methods suitable for the maintenance and growth ofmicrobial cultures are well known in the art. Techniques suitable foruse in the following examples may be found as set out in Manual ofMethods for General Bacteriology (Phillipp Gerhardt, R. G. E. Murray,Ralph N. Costilow, Eugene W. Nester, Willis A. Wood, Noel R. Krieg andG. Briggs Phillips, Eds.), American Society for Microbiology:Washington, D.C. (1994)); or in Manual of Industrial Microbiology andBiotechnology, 3^(rd) Edition (Richard H. Baltz, Julian E. Davies, andArnold L. Demain Eds.), ASM Press, Washington, D.C., 2010.

Production of Recombinant gtfJ in Fermentation

Production of the recombinant gtfJ enzyme in a fermenter was initiatedby expressing the gtfJ enzyme, constructed as described supra. A 10 mLaliquot of the seed medium was added into a 125 mL disposable baffledflask and was inoculated with a 1.0 mL culture of the E. coliMG1655/pMP52 prepared supra, in 20% glycerol. This culture was allowedto grow at 37° C. while shaking at 300 revolutions per minute (rpm) for3 hours.

A seed culture, for starting the fermenter, was prepared by charging a 2L shake flask with 0.5 L of the seed medium. 1.0 mL of the pre-seedculture was aseptically transferred into 0.5 L seed medium in the flaskand cultivated at 37° C. and 300 rpm for 5 hours. The seed culture wastransferred at optical density 550 nm (OD₅₅₀)>2 to a 14 L fermenter(Braun, Perth Amboy, N.J.) containing 8 L of the fermenter mediumdescribed above at 37° C.

Cells of E. coli MG1655/pMP52 were allowed to grow in the fermenter andglucose feed (50% w/w glucose solution containing 1% w/w MgSO₄.7H₂O) wasinitiated when glucose concentration in the medium decreased to 0.5 g/L.The feed was started at 0.36 grams feed per minute (g feed/min) andincreased progressively each hour to 0.42, 0.49, 0.57, 0.66, 0.77, 0.90,1.04, 1.21, 1.41 1.63, 1.92, 2.2 g feed/min respectively. The rate washeld constant afterwards by decreasing or temporarily stopping theglucose feed when glucose concentration exceeded 0.1 g/L. Glucoseconcentration in the medium was monitored using a YSI glucose analyzer(YSI, Yellow Springs, Ohio).

Induction of glucosyltransferase enzyme activity was initiated, whencells reached an OD₅₅₀ of 70, with the addition of 9 mL of 0.5 M IPTG(isopropyl β-D-1-thiogalacto-pyranoside). The dissolved oxygen (DO)concentration was controlled at 25% of air saturation. The DO wascontrolled first by impeller agitation rate (400 to 1200 rpm) and laterby aeration rate (2 to 10 standard liters per minute, slpm). The pH wascontrolled at 6.8. NH₄OH (14.5% weight/volume, w/v) and H₂SO₄ (20% w/v)were used for pH control. The back pressure was maintained at 0.5 bars.At various intervals (20, 25 and 30 hours), 5 mL of Suppressor 7153antifoam was added into the fermenter to suppress foaming. Cells wereharvested by centrifugation 8 hours post IPTG addition and were storedat −80° C. as a cell paste.

Preparation of gtfJ Crude Enzyme Extract from Cell Paste

The cell paste obtained above was suspended at 150 g/L in 50 mMpotassium phosphate buffer pH 7.2 to prepare a slurry. The slurry washomogenized at 12,000 psi (Rannie-type machine, APV-1000 or APV 16.56)and the homogenate chilled to 4° C. With moderately vigorous stirring,50 g of a floc solution (Aldrich no. 409138, 5% in 50 mM sodiumphosphate buffer pH 7.0) was added per liter of cell homogenate.Agitation was reduced to light stirring for 15 minutes. The cellhomogenate was then clarified by centrifugation at 4500 rpm for 3 hoursat 5-10° C. Supernatant, containing crude gtfJ enzyme extract, wasconcentrated (approximately 5×) with a 30 kilo Dalton (kDa) cut-offmembrane. The concentration of protein in the gftJ enzyme solution wasdetermined by the bicinchoninic acid (BCA) protein assay (Sigma Aldrich)to be 4-8 g/L.

Preparation of Polymer, Spinning Solutions, and Fiber Spinning Apparatusand Procedure

FIG. 1 is a schematic diagram of an apparatus suitable for use in thefiber spinning process hereof. The worm gear drive, 1, drives a ram, 2,at a controlled rate onto a piston fitted into a spinning cell, 3. Thespinning cell may contain filter assemblies. A suitable filter assemblyincludes 100 and 325 mesh stainless steel screens. A spin pack, 4,contains the spinneret, 5, and optionally stainless steel screens asprefilters for the spinneret. The extruded filament, 6, producedtherefrom is directed into a liquid coagulation bath, 7. In all theexamples listed in Table 1, the filament was extruded from the spinneretdirectly into the liquid coagulation bath—the bottom of the spinneretwas immersed in the bath.

The extrudate can be, but need not be, directed back and forth throughthe bath between guides, 8, which are normally fabricated of Teflon®PTFE. Only one pass through the bath is shown in FIG. 1. On exiting thecoagulation bath, 7, the thus quenched filament, 9, can optionally bedirected through a drawing zone using an independently driven roll, 10,around which the thus quenched filament is wrapped. The quenchedfilament may optionally be directed through a draw bath, 11, that allowsfurther treatment such as additional solvent extraction, washing ordrawing of the extruded filaments. The thus prepared filament is thendirected through a traversing mechanism, 12, to evenly distribute thefiber on the bobbin, and collected on plastic bobbins using a wind up,13. In one embodiment, the process comprises a plurality ofindependently driven rolls.

In one embodiment, the driven roll, 10, is removed from the fiberpathway, but the fiber is nevertheless immersed in the draw bath. Thetwo are independent of each other. In all of the examples, infra, thedriven roll, 10, was removed from the fiber pathway.

In one embodiment, a plurality of filaments is extruded through amulti-hole spinneret, and the filaments so produced are converged toform a yarn. In a further embodiment, the process further comprises aplurality of multi-hole spinnerets so that a plurality of yarns can beprepared simultaneously.

In each example, the wound bobbin of fiber produced was soaked overnightin a bucket of the liquid indicated in Table 1. The thus soaked bobbinof fiber was then air dried for at least 24 hours. The fiber tensileproperties were then determined according to ASTM D2101-82.

The spin cell, the piston, the connecting tubing and the spinneret wereall constructed of stainless steel.

Fiber Physical Property Measurement.

Physical properties such as tenacity, elongation and initial moduluswere measured using methods and instruments conforming to ASTM StandardD 2101-82, except that the test specimen length was 10 inches. Reportedresults are averages for 3 to 5 individual yarn tests.

The physical properties were determined for every fiber prepared. Theresults are shown in Table 1. Included are the denier of the fiberproduced, and the physical properties such as tenacity (T) in grams perdenier (gpd), elongation to break (E, %), and initial modulus (M) ingpd.

GLOSSARY OF TERMS

Column Label Actual Term Explanation Jet Vel. Jet Velocity The linearspeed of the fiber at the exit from (fpm) the spinneret. fpm Feet perminute Coag. Coagulation Temp. Temperature NA Not Applicable Theparameter does not apply to this example. NT Not Tested S.S.F. SpinStretch S.S.F. = (wind-up speed)/(jet vel.) Factor MeOH Methanol

Materials

Ingredient Stock No. Source Sucrose BDH8029 VWR Glucose G7528Sigma-Aldrich Dextran T-10 D9260 Sigma-Aldrich Boric Acid B6768Sigma-Aldrich NaOH SX0590-1 EMD

Example 1 Preparation of Polymer P1

Twenty liters of an aqueous solution was prepared by combining 1000 g ofsucrose (VWR #BDH8029), 20 g Dextran T-10 (Sigma #D9260), and 370.98 gBoric Acid (Sigma #B6768) were combined in about 18 l of water. 4N NaOH(EMD #SX0590-1) solution was employed to adjust the pH to 7.5.Additional water was then added to bring the total volume up to 20 l.The thus prepared solution was then charged with 180 ml of the enzymeextract prepared supra and allowed to stand at ambient temperature for48 hours. The resulting poly(α(1→3) glucan) solids were collected on aBüchner funnel using a 325 mesh screen over 40 micrometer filter paper.The filter cake was washed with deionized water and filtered as above.The deionized water wash was repeated three additional times. Finallytwo additional washes with methanol were carried out; the filter cakewas pressed out on the funnel and dried in vacuum at room temperature.Yield: 237.68 grams of white flaky solids.

Molecular weights were determined by size exclusion chromatography (SEC)with a GPCV/LS 2000™ (Waters Corporation, Milford, Mass.) chromatographequipped with two Zorbax PSM Bimodal-s silica columns (Agilent,Wilmington, Del.), using DMAc from J.T Baker, Phillipsburg, N.J. with3.0% LiCl (Aldrich, Milwaukee, Wis.) as the mobile phase. Samples weredissolved in DMAc with 5.0% LiCl. Number and weight average molecularweights (Mn and Mw) were found to be 139,000 and 279,000 Daltonsrespectively.

Example 1 Spinning Solution

A 250 mL wide mouth glass bottle was charged with 25 g of polymer P1 and225 g of 5 wt % sodium hydroxide. CS₂, (7.5 g), was then added via asyringe. The container was fitted with a cap and a septum through whicha polypropylene stirring rod had been fitted. The contents were manuallymixed with the stirring rod and then allowed to stand at roomtemperature overnight. The following day the partially dissolvedsolution (clear but containing a small amount of visible particulate)was transferred into a spin cell and piston containing screen packsincluding 100 and 325 mesh stainless steel screens. A piston was fittedover the viscous mixture. The mixture was then pumped back and forththrough 13 cycles using a motorized worm gear driven ram into anidentically equipped spinning cell coupled head to head with the firstcell via a coupler fabricated from ¼ inch stainless steel tubing.

Examples 2-4 Fiber Spinning

Approximately 20 hours after the preparation of the solution of Example1, the solution thus prepared was fed to the spinning apparatus, asdescribed, supra, referring to FIG. 1. The solution was fed to a 20 holespinneret wherein each hole was characterized by a circularcross-section, a diameter of 0.003 in and a length of 0.006 in. Table 1provides the spinning conditions that were used for the fibers preparedin Examples 2-4. The apparatus depicted in FIG. 1, as described supra,was modified by removal of the driven roll, 10, from the filamentpathway in Examples 1-3. The indicated spin stretch was attained byrunning the windup faster than the jet velocity. The solution of Example1 was metered at the rates shown in Table 1 through a spin pack having afilter assembly consisting of 100 and 325 mesh screens to the spinneret.The spinneret was immersed into a water coagulation bath containing, byweight, 8% H₂SO₄, 23% Na₂SO₄, and 0.5% ZnSO₄ The filament was extrudeddirectly into the quench bath vertically at the temperature indicated inTable 1. Additional residence time in the 6 foot long coagulation bathwas increased by directing the fiber over additional guide pins (8) fora total immersion distance of 4.1 or 11 ft. as indicated. In Examples 2and 3, upon removal from the coagulation bath the thus coagulatedfilaments was directed to a speed controlled wind-up with a traversingguide, at wind-up speeds shown in Table 1. In Example 4 the coagulatedfilaments were directed to a second bath of methanol for the length andat the temperature indicated in Table 1. In each example, severalhundred feel of fiber were wound onto a bobbin. Following wind-up, thefiber bobbins of Examples 2-4, were soaked sequentially respectively in5% NaHCO3, MeOH, and H₂O baths for a period of about 4 hours in each.The fiber was allowed to air dry before being subject to physicalmeasurements Physical properties were determined; results are shown inTable 1.

TABLE 1 JET COAGULATION/BATH SECOND BATH POLYMER % GLUCAN PUMP RATE VELLENGTH T LENGTH WIND-UP T E M EXAMPLE REF. SOLIDS (ml/min) (fpm)COMPOSITION (ft) (° C.) COMPOSITION (ft) T (° C.) SPEED (fpm) S.S.F.(gpd) (%) (gpd) DENIER 2 P1 9.7 1.60 56 8% H₂SO₄ 23% 11 51 NA NA NA 1262.3 1.0 3.7 60.2 40 Na₂SO₄ 0.5% ZnSO₄ 3 P1 9.7 1.60 56 8% H₂SO₄ 23% 1151 NA NA NA 100 2.3 1.2 4.9 60.5 75 Na₂SO₄ 0.5% ZnSO₄ 4 P1 9.7 1.60 568% H₂SO₄ 23% 4.1 51 MeOH 2.25 17 63 1.1 1.1 6.8 52.8 80.0 Na₂SO₄ 0.5%ZnSO₄ 6 P2 7.3 1.50 55 5% H₂SO₄ 3 21 NA NA NA 61 1.1 1.0 3.8 65.6 55 7P2 7.3 1.50 55 5% H₂SO₄ 3 21 MeOH 2.00 21 61 1.1 0.9 4.5 48.2 75 8 P27.3 1.50 55 5% H₂SO₄ 3 22 MeOH 2.00 23 82 1.5 1.1 4.2 64.6 47 9 P2 7.31.50 55 5% H₂SO₄ 3 24 MeOH 0.50 24 104 1.9 1.0 2.8 60.4 42 10 P2 7.31.50 55 Glacial 4.3 25 MeOH 1.80 26 57 1.0 1.2 5.4 70.4 60 Acetic 11 P27.3 1.50 55 50/50 Acetic acid/ 4.3 25 MeOH 1.80 26 56 1.0 1.0 2.5 62.770.0 H₂O v/v 13 P2 9.85 1.50 55 5% H₂SO₄ 4 24 NA NA NA 50 0.9 0.8 3.750.7 115 14 P2 9.85 1.50 55 5% H₂SO₄ 3.5 24 MeOH 1.15 25 60 1.1 1.3 4.771.4 75 15 P2 9.85 1.50 55 5% H₂SO₄ 3 24 MeOH 1.30 26 80 1.5 1.1 3.873.2 60 17 P3 7.33 2.10 75 5% H2SO4 4.3 12 NA NA NA 72 1.0 1.3 5.1 81 7518 P3 7.33 2.10 75 5% H2SO4 4.3 15 MeOH 1.5 17 72 1.0 N/A N/A N/A N/A 19P3 7.33 2.10 75 5% H2SO4 4.3 16 MeOH 1.5 16 89 1.2 N/A N/A N/A N/A 20 P37.33 1.60 50 10% H2SO4 4.3 17 NA NA NA 76 1.5 1.7 4.4 102 45 21 P3 7.331.60 50 10% H2SO4 4.3 18 NA NA NA 100 2.0 N/A N/A N/A N/A 22 P3 7.331.60 50 10% H2SO4 4.3 18 MeOH 1.9 13 62 1.2 N/A N/A N/A N/A 23 P3 7.331.60 50 10% H2SO4 4.3 19 H2O 1.66 45 33 0.7 1.8 5.3 85 115 25 P3 13 1.2845 10% H2SO4 4.2 24 NA NA NA 51 1.1 N/A N/A N/A N/A 26 P3 13 1.28 45 10%H2SO4 4.2 22 H2O 1.83 80 27 0.6 N/A N/A N/A N/A 27 P3 13 1.28 45 10%H2SO4 4.2 22 H2O 1.83 80 50 1.1 N/A N/A N/A N/A 28 P3 13 1.28 45 10%H2SO4 4.2 21 NA NA NA 72 1.6 N/A N/A N/A N/A 29 P3 13 3.20 110 10% H2SO44.2 20 NA NA NA 58 0.5 N/A N/A N/A N/A 30 P3 13 3.20 110 10% H2SO4 4.220 H2O 1.67 80 58 0.5 N/A N/A N/A N/A 31 P3 13 0.24 38 Glacial 4.2 24 NANA NA 28 0.7 N/A N/A N/A N/A Acetic NA = Not Applicable N/A = NotAvailable

Examples 5-11 Preparation of Polymer P2

Poly(α(1→3) glucan) polymer was synthesized, washed, and isolated usingthe materials and procedures employed for the preparation of Polymer P1in Example 1 except that 200 ml of the enzyme extract was added to thepH-adjusted sucrose/dextran/boric acid solution instead of 180 ml.Yield: 246.08 grams of white flaky solids.

Mn and Mw were determined as for polymer P1 to be 129,000 and 270,000respectively.

Example 5 Spinning Solution

A 250 mL wide mouth glass bottle was charged with 18 g of polymer P2 and225 g of 4.5 wt % sodium hydroxide. CS₂, (2.7 g), was then added viasyringe. The container was fitted with a cap and a septum through whicha polypropylene stirring rod had been fitted. The contents were manuallymixed with the stirring rod and then allowed to stand at roomtemperature overnight. After 72 hours the partially dissolved solutionwas transferred into a spin cell and piston containing screen packsincluding 325 mesh stainless steel screens. A piston was fitted over theviscous mixture. The mixture was then pumped back and forth through 13cycles using a motorized worm gear driven ram into an identicallyequipped spinning cell coupled head to head with the first cell via acoupler fabricated from ¼ inch stainless steel tubing.

Examples 6-11 Fiber Spinning

The fibers of Examples 6-11 were spun from the spinning solution ofExample 5, in the manner of the fibers of Examples 2-4, supra, under theconditions shown in Table 1. In Examples 6-9, the filament was extrudeddirectly into a coagulation bath containing 5% H₂SO₄ (aq.). In Example10, the fiber was extruded directly into a coagulation bath containingglacial acetic acid. In Example 11, into 50/50 acetic acid/water (v/v)Additional length in the coagulation bath was provided by directing thefiber over additional guide pins (8) for a total immersion distance of3, 4.3, or 4.5 ft. In Examples, 7-11, but not Example 6, upon removalfrom the coagulation bath the thus coagulated filament was directedthrough a second bath (11) of methanol at lengths and temperaturesindicated in Table 1. The fiber of Example 6 was guided directly to thewind-up. From the second bath, the coagulated fibers of Examples 7-11were directed to the wind-up, at the wind-up speeds shown in Table 1.The fiber bobbins were soaked as in Examples 2-4.

Physical properties were determined; results are shown in Table 1.

Examples 12-15 Example 12 Spinning Solution

A 250 mL wide mouth glass bottle was charged with 20 g of Polymer P2 and180 g of 4.5 wt % sodium hydroxide. CS₂, (3.0 g), was then added viasyringe. The container was fitted with a cap and septum through which apolypropylene stirring rod had been fitted. The contents were manuallymixed with the plastic stirrer and then allowed to stand 2 days. Thepartially dissolved solution was transferred into a 300 mL stainlesssteel cylinder fitted with 2×100 mesh, 1×325 mesh and 2×20 meshstainless steel screens. A piston was fitted over the viscous mixture.The mixture was then pumped back and forth through 13 cycles using amotorized worm gear driven ram into an identically equipped spinningcell coupled head to head with the first cell via a coupler fabricatedfrom ¼ inch stainless steel tubing.

Examples 13-15 Fiber Spinning

The fibers of Examples 13-15 were spun from the spinning solution ofExample 12 in the manner of the fibers of Examples 2-4, supra, under theconditions shown in Table 1. The fibers were extruded directly into 5%H₂SO₄ (aq.) at the temperature indicated in Table 1. The thus coagulatedfibers upon removal from the coagulation bath were directed to thewind-up at wind-up speeds shown in Table 1. The coagulated fibers ofExamples 14 and 15 were first passed through the second bath, asindicated in Table 1. The fiber bobbins were soaked and dried as inExamples 2-4.

Physical properties were determined; results are shown in Table 1.

Example 16 Preparation of Polymer P3

Poly(α(1→3) glucan) polymer was synthesized, washed, and isolated usingthe materials and procedures employed for the preparation of Polymer P1in Example 1 except that 200 ml of the enzyme extract was added to thepH-adjusted sucrose/dextran/boric acid solution instead of 180 ml.Yield: 228.52 grams of white flaky solids. M_(n) was 132,000 Daltons;M_(w) was 301,000 Daltons.

Example 16 Spinning Solution

A 250 mL wide mouth glass bottle was charged with 18 g of polymer P3 and225 g of 4.5 wt % sodium hydroxide. The container was fitted with a capand a septum through which a polypropylene stirring rod had been fitted.The contents were manually mixed with the stirring rod and then allowedto stand at room temperature overnight. CS₂, (5.4 g), was then added viasyringe the following morning. After the addition of CS₂ the partiallydissolved solution was immediately transferred into a spin cell andpiston containing screen packs including 325 mesh stainless steelscreens. A piston was fitted over the viscous mixture. The mixture wasthen pumped back and forth through 13 cycles using a motorized worm geardriven ram into an identically equipped spinning cell coupled head tohead with the first cell via a coupler fabricated from ¼ inch stainlesssteel tubing.

Examples 17-23 Fiber Spinning

Table 1 gives the spinning conditions that were used for the fibersprepared in Examples 17-23. The apparatus depicted in FIG. 1, asdescribed supra, was modified by removal of the driven roll, 10, fromthe filament pathway. Spin stretch was attained by running the windupfaster than the jet velocity. The spinning solution thus prepared wasmetered at the rates shown in Table 1 through a spin pack having afilter assembly consisting of 100 and 325 mesh stainless steel screensto a 20-hole spinneret having 0.003 inch diameter and 0.006 inch lengthholes. The filament was extruded directly into 5% H₂SO₄ for Examples17-19 and 10% H₂SO₄ for Examples 20-23 at the coagulation bathtemperature shown in Table 1. Upon removal from the coagulation bath thethus coagulated filament was directed through a second bath (11) ofmethanol at lengths and temperatures shown in Table 1, and thence to thewind-up. The filaments of Examples 17, 20, and 21 were guided directlyto the wind-up. The second bath in the case of Example 23 was filledwith water. Fiber spinning was completed within 8 hours from theaddition of carbon disulfide.

The fiber bobbins were soaked in 5% NaHCO₃ for 15 minutes, then soakedin water overnight. The bobbins were then removed and allowed to air drybefore being subjected to physical measurements.

Example 24 Spinning Solution

A 250 mL wide mouth glass bottle was charged with 32.9 g of polymer P3and 220 g of 5 wt % sodium hydroxide. The container was fitted with acap and a septum through which a polypropylene stirring rod had beenfitted. The contents were manually mixed with the stirring rod and thenallowed to stand at room temperature overnight. CS₂, (9.9 g), was thenadded via syringe the following morning. After the addition of CS₂ thepartially dissolved solution was immediately transferred into a spincell and piston containing screen packs including 325 mesh stainlesssteel screens. A piston was fitted over the viscous mixture. The mixturewas then pumped back and forth through 11 cycles using a motorized wormgear driven ram into an identically equipped spinning cell coupled headto head with the first cell via a coupler fabricated from ¼ inchstainless steel tubing.

Examples 25-31 Fiber Spinning

Table 1 gives the spinning conditions that were used for the fibersprepared in Examples 25-31. The apparatus depicted in FIG. 1, asdescribed supra, was modified by removal of the driven roll, 10, fromthe filament pathway for Examples 25-27. Spin stretch was attained byrunning the windup faster than the jet velocity. The spinning solutionthus prepared was metered at the rates shown in Table 1 through a spinpack having a filter assembly consisting of 100 and 325 mesh stainlesssteel screens to a 20-hole spinneret having 0.003 inch diameter and0.006 inch length holes. The filament was extruded directly into 10%H₂SO₄ for Examples 25-30 and glacial acetic acid for Example 31 at thecoagulation bath temperature shown in Table 1. Upon removal from thecoagulation bath the thus coagulated filaments of Examples 26, 27, and30 were directed through a second bath (11) of water at lengths andtemperatures shown in Table 1, and thence to the wind-up. The filamentsof Examples 25, 28, 29, and 31 were guided directly to the wind-up andwere not passed through the second bath. Fiber spinning was completedwithin 8 hours from the addition of carbon disulfide to the spinningsolution.

The fiber bobbins were soaked in 5% NaHCO₃ overnight, and then soaked inwater for an additional day. The bobbins were then removed and allowedto air dry before being subjected to physical measurements.

Examples 32-44, and Comparative Examples A-W

36 solutions were prepared to define the solution parameters thatresulted in solutions suitable for fiber spinning. For each of Examples32-44 and Comparative Examples A-W, 40 ml glass vials were charged withthe aqueous alkali metal hydroxide shown in Table 2. The concentrationof the alkali metal hydroxide solution, in weight-%, and the quantity ofthat alkali metal hydroxide solution are also shown in Table 2. 2 g ofPolymer P1 was then added to each vial. Carbon disulfide (CS₂) was addedin the amount shown in Table 2 and the vial was fitted with a septumthrough which a polypropylene stirring rod had been fitted. The contentswere manually mixed with the plastic stirrer and were allowed to standat room temperature for at least 12 hours with intermittent mixing. Thesolubility designations in Table 2 were determined by visual inspection.A clear solution was considered completely dissolved; a clear solutionwith some small particles floating around was also considered to bedissolved; since it was considered, that the partially dissolvedsolutions could be driven to complete dissolution with more intensivemixing. A turbid solution with large undissolved particles wasconsidered to be undissolved.

TABLE 2 NaOH Solution CS₂ Glucan Example # [NaOH] Weight (g) (g) Solids(%) Solubility 32 4.5 wt % 25.00 1.8 6.94 Sol 33 4.5 wt % 18.00 1.8 9.17Sol 34 4.5 wt % 14.75 1.8 10.78 Sol 35 4.5 wt % 25.00 0.6 7.25 Sol 364.5 wt % 18.00 0.6 9.71 Sol 37 4.5 wt % 14.75 0.6 11.53 Sol 38 4.5 wt %25.00 0.3 7.33 Sol 39 4.5 wt % 18.00 0.3 9.85 Sol 40 4.5 wt % 14.75 0.311.73 Sol 41 5 wt % 25.00 1.8 6.94 Sol Comp. Ex. A 5 wt % 18.00 1.8 9.17Ins Comp. Ex. B 5 wt % 14.75 1.8 10.78 Ins 42 5 wt % 25.00 0.6 7.25 SolComp. Ex. C 5 wt % 18.00 0.6 9.71 Ins Comp. Ex. D 5 wt % 14.75 0.6 11.53Ins 43 5 wt % 25.00 0.3 7.33 Sol 44 5 wt % 18.00 0.3 9.85 Sol Comp. Ex.E 5 wt % 14.75 0.3 11.73 Ins Comp. Ex. F 6 wt % 25.00 1.8 6.94 Ins Comp.Ex. G 6 wt % 18.00 1.8 9.17 Ins Comp. Ex. H 6 wt % 14.75 1.8 10.78 InsComp. Ex. I 6 wt % 25.00 0.6 7.25 Ins Comp. Ex. J 6 wt % 18.00 0.6 9.71Ins Comp. Ex. K 6 wt % 14.75 0.6 11.53 Ins Comp. Ex. L 6 wt % 25.00 0.37.33 Ins Comp. Ex. M 6 wt % 18.00 0.3 9.85 Ins Comp. Ex. N 6 wt % 14.750.3 11.73 Ins Comp. Ex. O 7.5 wt % 25.00 1.8 6.94 Ins Comp. Ex. P 7.5 wt% 18.00 1.8 9.17 Ins Comp. Ex. Q 7.5 wt % 14.75 1.8 10.78 Ins Comp. Ex.R 7.5 wt % 25.00 0.6 7.25 Ins Comp. Ex. S 7.5 wt % 18.00 0.6 9.71 InsComp. Ex. T 7.5 wt % 14.75 0.6 11.53 Ins Comp. Ex. U 7.5 wt % 25.00 0.37.33 Ins Comp. Ex. V 7.5 wt % 18.00 0.3 9.85 Ins Comp. Ex. W 7.5 wt %14.75 0.3 11.73 Ins

Example 45 Determination of Glucan Xanthate Formation and DecompositionUsing NMR Spectroscopy

2 g of poly(α(1→3) glucan) were dissolved in 25 ml of aqueous sodiumhydroxide (4.5 wt-%). After dissolution was complete, 0.6 g of carbondisulfide was added and the mixture so formed was then stirredmechanically and immediately transferred, using a syringe and needle,into a special 4.1 mm OD sample tube sold by New Era Enterprises, Inc.The tube was capped and lowered into a standard 7 inch, 5 mm NMR tubewhich contained 604 of D20 as an NMR lock solvent. These concentrictubes were placed in a small, bench-top centrifuge and spun for severalminutes to bring the sample to the bottom of the inner tube and toeliminate all air bubbles from the sample. The NMR tubes were removedfrom the centrifuge and placed in the magnet of a Bruker 500 MHz AvanceII Spectrometer equipped with a 5 mm CPDUL cryoprobe having z gradients.The probe was tuned and the magnet was shimmed before starting a seriesof consecutive experiments to investigate the formation and degradationof poly(α(1→3) glucan). Each experiment was acquired using the Brukerzgig pulse sequence with a spectral width of 33333.3 Hz (265.0 ppm), atransmitter offset of 160 ppm, and 32768 time domain points for anacquisition time of 0.4916 sec. A 3 sec. delay was used between pulsesand 3000 scans were acquired for each experiment, giving a total time of2 hours and 56 minutes for each experiment.

In order to suppress a baseline roll and obtain better peak integrals,the digital data from each experiment was converted to analog data sothat backward linear prediction could be performed. The first 12 pointsof each data set were replaced using Bruker's linear prediction dataprocessing based on the first 1024 data points. The Free Induction Decaywas also multiplied by a 2.0 Hz exponential function before beingtransformed. In order to determine the degree of xanthate substitutionthe integral area for the xanthate carbon, centered at 232.5 ppm, wascompared to the integral area (set to 1.00) for the glucan C1 anomericcarbons at 95.6-100.9 ppm, used as an internal calibration. At no pointin this series of experiments was a signal for the ¹³C of free CS₂(193.7 ppm) observed whereas signals for sodium trithiocarbonate (269.4ppm) and sodium carbonate (168.4 ppm) were present as by products of thedegradation of glucan xanthate over time. Results are shown in Table 3.

TABLE 3 Elapsed Time Degree of (hrs.) Xanthation 0 0.64 (est.) 3 0.61 60.52 12 0.43 24 0.23 36 0.15 54 0.04

1. (canceled)
 2. (canceled)
 3. (canceled)
 4. (canceled)
 5. (canceled) 6.(canceled)
 7. A process comprising forming a solution by dissolving in a0.75 to 2 molar aqueous alkali metal hydroxide, CS₂, and 5 to 15 percentby weight of the total weight of the resulting solution of poly(α(1→3)glucan) characterized by a number average molecular weight of at least10,000 Da, causing said solution to flow through a spinneret, forming afiber thereby; and contacting said fiber with an acidic liquidcoagulant; wherein said process the weight ratio of CS₂ to poly(α(1→3)glucan) lies in the range of 0.1 to 1.0.
 8. The process of claim 7wherein 7.5 to 15 percent by weight of poly(α(1→3) glucan) is dissolvedin said solution.
 9. The process of claim 7 wherein the alkali metalhydroxide is NaOH.
 10. The process of claim 7 wherein the concentrationof NaOH is 1.0 to 1.7 molar.
 11. The process of claim 7 wherein thepoly(α(1→3) glucan) 100% of the repeat units are glucose, and 100% ofthe linkages between repeat units are α(1→3) glycoside linkages.
 12. Theprocess of claim 7 further comprising allowing the solution to stand fora period of 1 to 8 hours prior to spinning.
 13. The process of claim 7wherein the poly(α(1→3) glucan) is characterized by a number averagemolecular weight in the range of 40,000-100,000 Daltons.
 14. The processof claim 7 wherein the CS₂ is added last.